Journal: IBRO Neuroscience Reports

Article Title: TGF-β1 modulates PFC glial cell activation to facilitate exercise-induced analgesia in mice with spared nerve injury

doi: 10.1016/j.ibneur.2026.03.009

Figure Lengend Snippet: Exercise modulates TGF-β1 expression in the prefrontal cortex (PFC) of mice 24 days after spared nerve injury (SNI). (a) Representative Western blot images of TGF-β receptor I (TGF-βR1) and TGF-β1 in the PFC. Tissue lysates from all experimental groups (SHAM, SHAME, SNI, SNIE) and recombinant human TGF-β1 (non-reduced and reduced) were loaded on the same SDS–PAGE gel, transferred to a single membrane, and probed with the same TGF-β1 antibody under identical exposure conditions. The recombinant protein (250 ng per lane) served as a positive control to verify the molecular weights of the dimeric (25 kDa) and monomeric (12.5 kDa) forms of TGF-β1. GAPDH was used as the loading control. (b-d) Quantitative Western blot analyses of (b) TGF-βR1, (c) dimeric TGF-β1 (25 kDa), and (d) monomeric TGF-β1 (12.5 kDa) expression levels in tissue lysates. Data are presented as mean ± SEM (n = 3). ** P < 0.01 vs. SHAM group; ## P < 0.01 vs. SNI group.

Article Snippet: To validate the specificity of the TGF-β1 antibody, Recombinant human TGF-β1 protein (Catalog # 240-B, R&D Systems, USA) was used as a positive control.

Techniques: Expressing, Western Blot, Recombinant, SDS Page, Membrane, Positive Control, Control

Journal: IBRO Neuroscience Reports

Article Title: TGF-β1 modulates PFC glial cell activation to facilitate exercise-induced analgesia in mice with spared nerve injury

doi: 10.1016/j.ibneur.2026.03.009

Figure Lengend Snippet: At 24 d after SNI, mouse PFC astrocytes were activated and microglia were unchanged. (a)Western blotting analysis of changes in GFAP and Iba1 expression in PFC (n = 3); (b) Quantification of GFAP in PFC; (c) Quantification of Iba1 in PFC; (d) MFI representative images of GFAP in PFC; (e) MFI representative image of Iba1 in PFC; (f) Quantification of GFAP in PFC. Values represent mean ± SEM (Scale bar =75μm, 9 PFC sections from 3 mice per group); (g) Quantification of Iba1 in PFC. Values represent mean ±SEM (Scale bar = 75μm, 9 PFC sections from 3 mice per group). Values represent the mean ±SEM. * P < 0.05, ** P < 0.01, compared with SHAM group; # P < 0.05, ## P < 0.01, compared with SNI group, the difference was statistically significant; (h) Representative MFI images of changes in the colocalization of TGF-β1(red) and astrocytes (green) in the PFC; (i) Quantification of TGF-β1 and astrocytes in PFC. Values represent the mean ± SEM (Scale bar =100μm, nine PFC sections from three mice per group). * P < 0.05 versus the SHAM group; # P < 0.05 versus the SNI group.

Article Snippet: To validate the specificity of the TGF-β1 antibody, Recombinant human TGF-β1 protein (Catalog # 240-B, R&D Systems, USA) was used as a positive control.

Techniques: Western Blot, Expressing

Journal: IBRO Neuroscience Reports

Article Title: TGF-β1 modulates PFC glial cell activation to facilitate exercise-induced analgesia in mice with spared nerve injury

doi: 10.1016/j.ibneur.2026.03.009

Figure Lengend Snippet: TGF-βRI inhibition reverses exercise-induced analgesia and modulates glial activation in the PFC. (a, b) Time course of mechanical and cold hyperalgesia tests (n = 9). The green shading indicates the duration of the exercise intervention, and the green vertical lines denote the timing of intrathecal injections. Data are presented as mean ± SEM. ** P < 0.01 versus the SNIE group, # P < 0.05, ## P < 0.01 vs. SC group. (c) Representative Western blot images of TGF-βR1 and TGF-β1 in the PFC. Tissue lysates from SC and SA groups and recombinant human TGF-β1 (100 ng per lane) (non-reduced and reduced) were loaded on the same SDS–PAGE gel, transferred to a single membrane, and probed with the same TGF-β1 antibody in a single exposure without splicing. The recombinant protein served as a positive control to verify the molecular weights of the dimeric (25 kDa) and monomeric (12.5 kDa) forms of TGF-β1. GAPDH was used as the loading control. (d-f) Quantitative analysis of (d) TGF-βR1, (e) dimeric TGF-β1 (25 kDa), and (f) monomeric TGF-β1 (12.5 kDa) expression levels (n = 3). (g-i) Western blot analysis of glial markers. (g) Representative images of GFAP and Iba1 with GAPDH control. Quantitative analysis of (h) GFAP and (i) Iba1 expression levels (n = 3). (j, k) Representative immunofluorescence images showing the expression of (j) GFAP and (k) Iba1 in the PFC. Scale bar = 75 μm. (l, m) Quantification of the mean fluorescence intensity (MFI) for (l) GFAP and (m) Iba1 (n = 9 sections from 3 mice per group). Data in bar graphs are presented as mean ± SEM. * P < 0.05, ** P < 0.01 vs. SC group. SC: Spared nerve injury with exercise training followed by intrathecal (i.t.) injection of saline; SA: Spared nerve injury with exercise training followed by i.t. injection of the TGF-βRI inhibitor.

Article Snippet: To validate the specificity of the TGF-β1 antibody, Recombinant human TGF-β1 protein (Catalog # 240-B, R&D Systems, USA) was used as a positive control.

Techniques: Inhibition, Activation Assay, Western Blot, Recombinant, SDS Page, Membrane, Positive Control, Control, Expressing, Immunofluorescence, Fluorescence, Injection, Saline

Journal: bioRxiv

Article Title: Partial EMT Drives Persistent Collective Migration via Collision Guidance in Heterogeneous Populations

doi: 10.64898/2026.04.07.714519

Figure Lengend Snippet: (a) Experimental design compares cells in untreated control with 7 day treatment with 1 ng ml −1 or 5 ng ml −1 TGF- β 1. Cells were then replated for live imaging. (b) Representative snapshots of cell velocity fields for all three conditions (green arrows). Cell cytoplasm is labeled in red (CellTracker Deep Red) and nuclei in blue (Hoechst). (c) Comparison of average cell velocity and correlation length for each experimental condition from 15-22.5 h. Elliptical contours illustrate the Gaussian summaries of the data distribution, constructed from the mean and covariance matrix. (d) Schematic and representative images for single cell trajectories reconstructed from optical flow. (e) Representative cell trajectories classified as slow and persistent, slow and random, fast and persistent, or fast and random. (f) Comparison of representative single cell trajectories for control relative to TGF- β 1 treatments, revealing distinct behaviors at early and later times.

Article Snippet: EMT was induced in MCF-10A cells using 1 ng/mL or 5 ng/mL TGF- β 1 (R&D Systems, 240-B002) in growth media.

Techniques: Control, Imaging, Labeling, Comparison, Construct, Single Cell

Journal: bioRxiv

Article Title: Partial EMT Drives Persistent Collective Migration via Collision Guidance in Heterogeneous Populations

doi: 10.64898/2026.04.07.714519

Figure Lengend Snippet: (a) Representative cell trajectory, partitioned into three sequential time segments: T seg (i), 00:00–07:30; T seg (ii), 07:30–15:00; and T seg (iii), 15:00–22:30. (b) UMAP dimensionality reduction of 17 motility metrics for each 7.5 h segment (per trajectory) projected into two dimensions, and overlaid with illustrative single cell trajectory segments. UMAP1 correlates with fast to slow migration, while UMAP2 correlates with persistent to random migration. (c) Spearman correlation between UMAP components and 17 individual motility features. (d, e) Distribution of cell trajectory segment metrics from each experimental condition at time segments T seg (i) and T seg (iii) (Control, brown; TGF- β 1 1 ng ml −1 , gray; and TGF- β 1 5 ng ml −1 , mint). Kernel density distributions of each condition along the UMAP1 and UMAP2 components are shown in the top and right panels, respectively. (f) Centroids for each time segment and experimental condition in the UMAP projection. Arrows indicate the displacement of centroids between adjacent segments, revealing changes in motility behavior over time. (g) Pairwise overlap analysis indicating relative similarity between experimental conditions and time windows. Smaller values indicate greater differences in motility behavior. Temporal changes in representative motility features across time segments: (h) speed, (i) directionality, (j) velocity correlation, and (k) arrest coefficient. Each dot represents the mean value of the corresponding motility feature, and error bars indicate the standard error of the mean (SEM).

Article Snippet: EMT was induced in MCF-10A cells using 1 ng/mL or 5 ng/mL TGF- β 1 (R&D Systems, 240-B002) in growth media.

Techniques: Single Cell, Migration, Control

Journal: bioRxiv

Article Title: Partial EMT Drives Persistent Collective Migration via Collision Guidance in Heterogeneous Populations

doi: 10.64898/2026.04.07.714519

Figure Lengend Snippet: (a) Representative images of MCF-10A expressing Z-cad dual fluorescent reporter in untreated control relative to 1 ng/mL and 5 ng/mL TGF- β 1 treatments. (b) Distribution of EMT reporter color expression at 0, 9, and 18 hours after imaging in control vs TGF- β 1 treatment. (c) Representative cell morphologies defined as compact, elongated, or spread. (d, e) Distribution of cell color and morphology in the UMAP latent space at different time segments T seg (i);(d), T seg (iii);(e). Cell shapes are reconstructed from autoencoder-derived latent representations, while color indicates the average EMT reporter level computed over local grid regions in the UMAP space. Subpanels summarize color distributions and representative morphological features. (f) Pairwise overlap analysis indicating relative similarity in motility behavior between EMT reporter and experimental condition. (g, h) Centroids for each time segment for fluorescent reporter state and experimental condition. Arrows indicate the displacement of centroids between adjacent segments, revealing changes in motility behavior over time.

Article Snippet: EMT was induced in MCF-10A cells using 1 ng/mL or 5 ng/mL TGF- β 1 (R&D Systems, 240-B002) in growth media.

Techniques: Expressing, Control, Imaging, Derivative Assay

Journal: bioRxiv

Article Title: Partial EMT Drives Persistent Collective Migration via Collision Guidance in Heterogeneous Populations

doi: 10.64898/2026.04.07.714519

Figure Lengend Snippet: (a) Ternary diagram showing the percent composition of fluorecent reporter expression(red, green, and uncolored) across experimental conditions. Orange circles: Control; gray squares: TGF- β 1 (1 ng mL −1 ); cyan triangles: TGF- β 1 (5 ng mL −1 ). (b) Heterogeneity index is a read out of how evenly the three EMT states are distributed, where 0 corresponds to equal percentages of each EMT state in the population and 1 corresponds a population wholly comprised of one EMT state. Box plots show median and interquartile range with overlaid individual data points. Pairwise comparisons were evaluated using Bonferroni-corrected post hoc tests (*** p < 0.001). (c) Representative images of local color heterogeneity within a small region of interest. (d) Color heterogeneity in control or TGF- β 1 treatment condition. Horizontal lines indicate the median and interquartile range (IQR); dots denote individual cells. Statistical significance was assessed using Bonferroni-corrected pairwise comparisons (*** p < 0.001). Density–heterogeneity maps of cell motility metrics; (e) speed and (f) velocity correlation. Data were binned along number of cells and color heterogeneity of each patch. Overlaid points represent individual measurements.

Article Snippet: EMT was induced in MCF-10A cells using 1 ng/mL or 5 ng/mL TGF- β 1 (R&D Systems, 240-B002) in growth media.

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: Partial EMT Drives Persistent Collective Migration via Collision Guidance in Heterogeneous Populations

doi: 10.64898/2026.04.07.714519

Figure Lengend Snippet: (a) Schematic illustration of collective collision dynamics and experimental setup using a PDMS stencil to induce controlled collective collisions. (b, c) Mean migration speed of expanding direction and correlation length of expanding monolayer across TGF- β 1 pretreatment conditions ( n = 3 independent experiments). Statistical comparisons were performed using one-way ANOVA, followed by Welch’s unequal-variance t -tests with Bonferroni correction for multiple comparisons. Significance levels are indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. (d) Representative snapshots of monolayer expansion and collisions between distinct epithelial or EMT states. Kymograph of collective front and cell velocity for control-control collisions (e,f), TGF- β 1 (1 ng mL −1 )–control collision (g,h), and TGF- β 1 (1 ng mL −1 )–TGF- β 1 (5 ng mL −1 ) collision (i,j). Orange stars indicate the spatiotemporal position of collective collisions (k) Schematic of collision behaviors associated with arrest, deformation wave, and repulsion.

Article Snippet: EMT was induced in MCF-10A cells using 1 ng/mL or 5 ng/mL TGF- β 1 (R&D Systems, 240-B002) in growth media.

Techniques: Migration, Control